1.Differential expression profile of microRNAs in different types of uveal melanoma
Yinan, LIU ; Lei, SHAO ; Wenbin, WEI
Chinese Journal of Experimental Ophthalmology 2017;35(9):778-785
Background Researches showed that microRNA (miRNA) is involved in the pathogenesis and development of many tumors and plays a cancer-suppressing-gene like role or cancer-gene like action.Uveal melanoma (UM) is a common ocular malignant tumor in aduh,and the mechanism of UM pathogenesis and metastasis is still not elucidated.Understanding the differential expression of miRNAs in UM is expected to provide a basis for targeting treatment of UM.Objective This study was to screen and compare the expression profiles of miRNAs in epithelial type and spindle type of UM.Methods The use of specimens of UM and donor eyes was approved by Ethic Commission of Capital Medical University.The specimens of epithelial type (4 specimens) and spindle type (4 specimens) of UM confirmed by histopathology and immunochemistry were collected in Beijing Tongren Hospital from March 2013 to October 2015.The expression profile of miRNA was assayed by miRNA array.Normal uveal specimens were obtained from 8 donors as controls.The differentially expressing miRNAs were screened by intergroup differential folds of ≥2.The genes targeting differentially expressed miRNAs were predicted using multiples online software and the potential signal pathway was further analyzed by bioinformatics method.The microarray outcomes were validated by real-time quantitative PCR.Results Spindle cell type and epithelial cell type of UMs were verified by hematoxylin and eosin staining.Immunochemistry showed that HMB45,melanin-A and S-100 were positively expressed in the two types of UM.Compared with the normal uveal tissue,109 differentially expressed miRNAs,including 29 up-regulated and 80 down-regulated miRNAs were seen in the spindle cell type of UM,and in the epithelial cell type of UM,50 differentially expressed miRNAs were found,including 23 up-regulated and 27 down-regulated miRNAs.In spindle cell type of UM,the up-regulated miRNAs were miR-146a-5p,miR-25-3p and miR-29b-l-5p,and down-regulated ones were miR-126-5p,miR-183-5p and miR-96-5p.In epithelial cell type of UM,the up-regulated miRNAs were miR-155-5p,miR-210 and miR-378a-5p,and down-regulated ones were miR-199a-5p,miR-143-3p and miR-143-5p.In addition,the mutual up-regulated miRNA in both spindle cell type of UM and epithelial cell type of UM were miR-132-3p,miR-21-5p,miR-34a-5p and miR-34b-5p,and mutual down-regulated ones were miR-125b-2-3p,miR-126-3p,miR-199a-3p and miR-214-3p.Bioinformatics analysis showed that the targeting genes predicted by differentially expressed miRNAs participated in a number of biological pathways,including cancer-related pathway,mitogenactivated protein kinase (MAPK) pathway,Wnt signal pathway and intercellular adhesion,endocytosis,prostatic cancer,colorectal cancer pathways.Conclusions Many differentially expressed miRNAs exist among spindle cell type of UM,epithelial cell type of UM and normal uveal tissue.These miRNAs participate in or regulate the biological behaviour of UM via different signal pathways.
2.Analysis of diarrhea in 68 patients subsequent to orthotopic liver transplantation
Weidong DUAN ; Lei HE ; Wenbin JI
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
Objective To investigate the factors in association with colorectal disorders in adult recipients of liver allograft. Methods A retrospective cohort study was carried out with clinical, microbiological and management data regarding diarrhea in 218 adult recipients of liver allograft from Jan. 2003 to Dec. 2006. Results Of the 218 patients, 68(31.2%) of them, who did not history of ulcerative colitis, were found to have diarrhea after orthotopic liver transplantation. Among these 68 patients, diarrhea was probably caused by administration of immunosuppressive agents in 35 cases (51.4%), in 12 cases (17.6%) diarrhea was antibiotics associated, and in 5 cases (7.4%) it was due to intraperitoneal infection. The other pathogenic factors included fungal infection in 3 cases (4.4%), cytomegalovirus infection in 3 cases (4.4%), Roux-en-Y choledochojejunostomy in 8 cases (11.8%) and some other unknown problem related in 2 cases (2.9%). Diarrhea occurred soon after transplantation in most cases. Of the 68 patients, the symptom of diarrhea occurred in the first 2 months in 45 cases (66.2%), and in the next 4 months in 23 cases (33.8%), i.e. the symptoms occurred 5-180 days after transplantation. Conclusion Immunosuppressive agents, antibiotics, fungal infection and cytomegalovirus infection are the top four common causes of diarrhea after orthotopic liver transplantation. The outcome is good with appropriate conservative management.
3.Effect of morin on LPS induced acute lung injury and its mechanism
Jie WANG ; Yingkai FENG ; Wenbin ZHANG ; Jie MOU ; Wenhui LEI
Chongqing Medicine 2015;(19):2609-2612
Objective To study the effect of morin on LPS induced acute lung injury mouse model and its mechanism .Meth‐ods Thirty male C57B/L mice were randomly divided into control group ,LPS group and LPS+ morin group ,with 10 in each group .5 mg/kg LPS was instilled into the lung from an trachea intubation in LPS group and LPS+morin group .Then the mice in LPS+morin group received an intraperitoneal injection of morin (40 mg/kg) every day for the next 3 d .Others received an equal a‐mount of saline .After 72 h ,the mice were sacrificed .The bronchoalveolar lavage fluid (BALF) was collected and centrifuged;the sediments were stained with Wright‐Giemsa for total cell and neutrophil count and the supernates were prepared for ELISA .The wet and dry weight of lung was weighed to calculate the wet/dry weight ratio .HE staining was performed to examine the pathologi‐cal change of lung .Western blot was used to determined the expression of TLR4 ,IKK and NF‐κB .Results Intratracheal instillation of LPS successfully established ALI model in mouse .LPS caused significant pathological changes including inflammatory cells infil‐tration ,alveolar septa thickness ,hemorrhage and edema .The wet/dry weight ratio ,the total cell count ,neutrophil count ,TNF and IL‐1βlevel in BALF ,and the expression of TLR4 ,NF‐κB ,and IKK were all increased significantly (P<0 .05) ,which were allevia‐ted by intraperitoneal injection of morin .Conclusion Morin can dampen the inflammatory response during LPS induced ALI in mouse ,which is potentially attributed to its inhibitory effect on the activation of NF‐κB .
4.Three-dimensional finite element simulation and biomechanical analysis of human mandible injury
Tao LEI ; Wenbin TU ; Tingting WANG ; Jiajun ZHENG ; Liangxian XIE
Chongqing Medicine 2016;(2):151-155
Objective The simulation of the human mandible injury was carried out by using the finite element simulation technology ,and the biomechanical analysis of simulation results was developed to explore the mechanism of injuries .Methods The Chinese Visible Human digital data were used to establish the three-dimensional element model of mandible injuries ,and the dynam-ic processes of human mandible injuries in different conditions were simulated ,and the biomechanical analysis were carried out by u-sing the Von Mises stress and effective strain .Results The three-dimensional element model of mandible injuries was established , the dynamic damage and fracture of human mandible were simulated successfully ,the mandibular angle and condylar were the predi-lection parts of high-stress ,high-strain and fractures .Conclusion The Von Mises stress and effective strain can be used to predict and judge the bone tissue injuries ,the finite element method can simulate the impact injuries of mandible effectively ,and the simula-ted results can provide guidance and reference for basic research and clinical treatment of oral and maxillofacial injuries .
5.A correlated study between-L-alpha glucosidase and intrauterine growth and glucose and lipid metabolism in fetus below the gestational age
Wenbin DONG ; Xiaopin LEI ; Lingpin ZHANG ; Lianyu ZHANG ; Na HE
The Journal of Practical Medicine 2017;33(10):1605-1607
Objective To investigate the change of serum alpha-L-fucosidase(AFU)and its correlation with the blood glucose and lipid level in small for gestational age(SGA)fetuses. Methods 125 SGA fetuses and 128 fetuses in appropriate for gestational age(AGA)with wet lung were treated in our hospital and were investigated as case control study. The serum of AFU ,blood glucose and lipid were measured and compared within 24 hours after birth in these 2 groups. Results Comparing with the AGA infants ,the SGA babies had lower level of serum AFU,high density lipoprotein,apolipoprotein A and apolipoprotein B(P<0.05). The correlation analysis showed that the serum AFU level has positive correlation with blood glucose,total cholesterol,high density lipoprotein, low density lipoproteinand apolipoprotein A(P < 0.05),while it has negative correlation with serum triglycerides in SGA(P<0.05). Conclusions The SGA infants have lower level of serum AFU and lipid metabolic disorders after birth,and its serum AFU level has correlation with its blood glucose and lipid level.
6.Effect of promethazine on eIectrophysioIogicaI action of guinea pig hearts
Wenbin SHANG ; Zhuoka LUO ; Xuehua LL ; Lei LLU ; Wei WANG ; Kesu CHEN ; Zhongyue WANG ; Long CHEN
Chinese Journal of Pharmacology and Toxicology 2014;(5):691-696
OBJECTIVE To explore the effect and underlying mechanism of promethazine(PMZ) on proarrhythmia in guinea pigs. METHODS ① InvivoECG recordings were made to analyze effects of jugular intravenous(iv)injection of PMZ on ECG in guinea pigs. PMZ was injected in this order:3.83→7.67→15.33→38.33 mg·kg-1 cumulatively. ② In vitroECG recordings were made to analyze effects of PMZ on ECG in isolated hearts of guinea pigs. PMZ was perfused in such order:0. 1 → 1 → 10 →50 μmol·L-1 . ③ L-type Ca2+ currents from ventricular myocytes in guinea pigs were recorded to investi-gate the PMZ's blocking effect. PMZ was perfused in such order:0.1→1→10→50 μmol·L-1→washout.④ hNav1.5 and hERG currents were recorded to investigate the PMZ's blocking effects. PMZ-perfused in such order:1→3→10→30 μmol·L-1 for hNav1.5 current analysis,and 0.3→1→3→10 μmol·L-1 for hERG current analysis. RESULTS ① PMZ(15.33 mg·kg-1 )significantly prolonged QRS intervals in guinea pigs invivoECG(P﹤0.05). PMZ(38.33 mg·kg-1 )prolonged QRS,QTc,and P-R intervals but reduced the heart rate( P﹤0.05). PMZ(10 μmol·L-1 )reduced the heart rate of isolated guinea pig hearts. PMZ 50 μmol·L-1 prolonged QRS and QTc intervals and further reduced the heart rate(P﹤0.05).③ PMZ inhibited the L-type Ca2+ current from ventricular myocytes in guinea pigs in a concentration-dependent manner with the lC50 of(8.9±1.0)μmol·L-1 . ④ PMZ inhibited the hNav1.5 and hERG currents in a concentration-dependent manner with the lC50 of 6.1±1.5 and(1.6±0.2)μmol·L-1 ,respectively. CONCLUSION PMZ might cause arrhythmia at overdoses and incombination with other drugs which have potential blocking effect on /Na ,Ca2+ and /kr currents. The proarrhythmic effect of PMZ might be mediated by the blocking effect on /Na ,Ca2+ and /kr currents.
7.The role of PKCβ/p66Shc oxidative stress signaling pathway in hyperoxia induced apoptosis in alveolar epithelial cell A549
Zhongli CHE ; Wenbin DONG ; Qingping LI ; Xiaoping LEI ; Lan KANG ; Lin GUO ; Xuesong ZHAI ; Feng CHEN
Journal of Clinical Pediatrics 2013;(11):1066-1069
Objectives To explore the role of PKCβ/p66Shc oxidative stress signaling pathway in hyperoxia-induced apoptosis of alveolar epithelial cells A549. Methods A549 cells were cultured in vitro and divided randomly into control (incubated with 5%CO2), hyperoxia group (exposed to a mixture of 900 ml/L O2 and 50 ml/L CO2 at speed of 3 L/min for 10 mins, then cultured in a closed environment) and LY333531 group (treated with 10μmol/L of PKCβinhibitor LY333531 for 24h then induced with hyperoxia for 10 mins). The cellular morphology was observed under inverted microscope at 12, 24 and 48 h of treatment. The cell apoptosis was detected by lfow cytometry. Expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 were detected by immunohistochemistry after 24 h of treatment. Results Comparing to the control group, the cellular morphology of A549 in the hyperoxia group changed to spherical shapes and space between cells increased, the living cell count decreased and suspension cell increased. The living cell count in LY333531 group increased and suspension cell decreased than those in hyperoxia group but not reach the levels of the control group. The apoptosis rate of A549 cells and the expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 at 24 h were signiifcantly increased in the hyperoxia group than those in the control group, while the apoptosis rate and the expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 were greatly decreased in the LY333531 group than those in the hyperoxia group (all P<0.01). Conclusions The expression of PKCβin A549 cells can be increased by the hyper-oxia induction but reduced by LY333531, and then the expressions of Pin1, p66Shc and p66Shc-Ser36 are reduced. Thus the re-duced apoptosis of A549 cells relieve the cell injury induced by hyperoxia.
8.Antidepressant effect of total flavonoids extracted from Xiaobuxin-Tang in forced swimming tests and learned helplessness in rats and mice
Youzhi ZHANG ; Nengjiang YU ; Li YUAN ; Lei AN ; Yimin ZHAO ; Wenbin XIAO ; Zhipu LUO ; Yunfeng LI
Chinese Journal of Pharmacology and Toxicology 2008;22(1):1-8
AIM Xiaobuxin-Tang (XBXT) is a traditional Chinese herbal decoction which is composed of Haematitum, Flos Inulae, Folium Phyllostachydis Henonis and Semen Sojae Preparatum. The present study was to investigate if the total flavonoids extracted from XBXT (XBXT-2) had antidepressant effect. METHODS Forced swimming tests in mice and rats, and learned helplessness (LH) model of rats were adopted to affirm the antidepressant effect of XBXT-2 with the test on spontaneous motor activity. Plasma corticosterone level in the LH rats was measured with ELISA. RESULTS Single administraton of XBXT-2 at the doses of 50 and 100 mg·kg-1 (ig) significantly decreased the duration of immobility time in the forced swimming tests in mice and rats. Researches on LH model of rats indicated that XBXT-2 at doses of 50 and 25 mg·kg-1 markedly reduced the number of escape failure in shuttle box. Meanwhile, the plasma corticosterone level of the LH rats was significantly decreased. XBXT-2 50-200 mg·kg-1 had no effects on spontaneous motor activity in mice. CONCLUSION XBXT-2 possesses significant antidepressant-like effect. The mechanism may involve the inhibition of the hyperaction of the hypothalamic-pituitary-adrenal axis.
9.Screening of tumor mark for the lung cancer by using peptide ZS-6
Wenbin FENG ; Xuediao PAN ; Jie ZHOU ; Lei SHI ; Junqiang LUO ; Xun LI ; Linquan ZANG
Chinese Pharmacological Bulletin 2010;26(1):44-47
Aim To select a potential biomarker for early lung cancer diagnosis and targeted therapy by using the cancer specific bounded peptide ZS-6 which had already been obtained from the laboratory.Methods The peptide ZS-6 marked by biotin was used as a probe to pan out the human lung cancer cDNA phage display library,after 4 rounds of subtraction panning,the specific binding clones of ZS-6 were identified.After amplification and purification,then those DNA sequences were identified and analyzed with bioinformatics.Results 18 phage clones were identified to the specific peptide ZS-6 and the DNA sequence showed one of them was an unknown new gene while the others were known tumor related genes.Conclusion A tumor biomarker selected from human lung cancer cDNA library provides a potential tool for early lung cancer diagnosis and therapy.
10.Role and mechanism of Astragalus in alleviating adhesion of neutrophils to human renal tubular cells induced by postasphyxial-serum of neonate
Hongyu YANG ; Wenbin DONG ; Qingping LI ; Xiaoping LEI ; Tao XIONG ; Cunliang DENG ; Feng CHEN
Journal of Clinical Pediatrics 2009;(11):1070-1073
Objective To investigate the role of Astragahs in alleviating adhesion of neutrophils to HK-2 cells induced by postasphyxial-serum of neonate and its signal transduction mechanism. MethodsHK-2 cells were used as target cell. Control group, asphyxia group, Astragalus group were divided in the experiment. The 20% (volume fraction) postasphyxial-serum was used as attacking factor. The following indicators were detected: cellular morphology and neutrophils adhesion were observed with inverted microscope. The activity of myeloperoxidase (MPO) in the cell suspension were determined by biochemistry assay as the indicator for adhesion of neutrophils to HK-2. Intercellular adhesion molecule-1(ICAM-1) were examined by flow eytometer. ResultsThe neutrophils numbers (MPO activity) and ICAM-1 expression in asphyxia group were higher than that of the control group, but dramatically decreased in Astragalus group (P < 0.05). ConclusionsThese data demonstrated that Astragalus could alleviate the adhesion of neutrophils from neonate with asphyxia to HK-2 and it' s intracellular signal transduction mechanism is presumably involved in the inhibition of ICAM-1 expression on HK-2 cellular membrane.